US8999364B2 - Implantable article, method of forming same and method for reducing thrombogenicity - Google Patents
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- US8999364B2 US8999364B2 US11/753,896 US75389607A US8999364B2 US 8999364 B2 US8999364 B2 US 8999364B2 US 75389607 A US75389607 A US 75389607A US 8999364 B2 US8999364 B2 US 8999364B2
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/0077—Special surfaces of prostheses, e.g. for improving ingrowth
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
- A61F2/06—Blood vessels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
- A61L29/08—Materials for coatings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
- A61L31/10—Macromolecular materials
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/0005—Use of materials characterised by their function or physical properties
- A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L67/00—Compositions of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Compositions of derivatives of such polymers
- C08L67/04—Polyesters derived from hydroxycarboxylic acids, e.g. lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2210/00—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2210/0004—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof bioabsorbable
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/06—Coatings containing a mixture of two or more compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/08—Coatings comprising two or more layers
Definitions
- the present invention relates generally to implantable devices, such as implantable medical devices, and methods for the manufacture thereof.
- the invention also relates to methods for enhancing and promoting endothelialization and for minimizing thrombus formation on the surface of the implantable device.
- Thrombus formation is a very complex process involving inter-dependent interactions between a surface of an implantable device, platelets and coagulation proteins.
- the present invention addresses the problem of thrombosis by endothelialization of a bodily fluid or tissue-contacting, particularly blood-contacting, surface to render that surface substantially non-thrombogenic.
- the invention also addresses the problem of thrombosis by providing an eroding layer on the surface that results in the removal of any thrombus formation as the layer erodes.
- an implantable device having at least one surface for contacting bodily fluid or tissue, said at least one surface comprising a plurality of nano-craters thereon that enhance or promote endothelialization of said at last one surface.
- an implantable device having at least one first degradable layer providing at least one surface of the implantable device for contacting bodily fluid or tissue and disposed about a central core, and at least one second degradable layer between said first degradable layer and the central core, wherein said first degradable layer has a first degradation rate and said second degradable layer has a second degradation rate such that said at least one first degradable layer degrades more rapidly than said at least one second degradable layer on contact with bodily fluid or tissue.
- the material of the implantable device is not particularly limited.
- the nano-craters may be formed in the material that constitutes the body of the implantable device, or may be formed in a layer that is applied to a support substrate forming the implantable device.
- the nano-craters will be formed in a surface layer of suitable biocompatible material applied to a support structure for the implantable device.
- the options for the biocompatible material forming the outer layer of the implantable device are generally known and are discussed hereafter.
- the form of the implantable device is similarly not particularly limited. This may include any device that is intended to come into contact with bodily fluids or tissues, be that during in vivo applications or in vitro applications. Examples of particular devices will be provided hereafter.
- an implantable device having at least one surface for contacting bodily fluid or tissue comprising: providing on said at least one surface a plurality of nano-craters that enhance or promote endothelialization of said at least one surface.
- a method of reducing thrombogenicity of an implantable device having at least one surface for contacting bodily fluid or tissue, or promoting or enhancing endothelialization of an implantable device having at least one surface for contacting bodily fluid or tissue comprising: providing on said at least one surface a plurality of nano-craters that enhance or promote endothelialization of said at least one surface.
- a method of manufacturing an implantable device having at least one surface for contacting bodily fluid or tissue comprising: providing at least one first degradable layer which provides said at least one surface and which is disposed about a central core, and at least one second degradable layer between said first degradable layer and the central core, wherein said first degradable layer has a first degradation rate and second degradable layer has a second degradation rate such that said at least one first degradable layer degrades more rapidly than said at least one second degradable layer on contact with bodily fluid or tissue.
- a method of reducing thrombogenicity of an implantable device having at least one surface for contacting bodily fluid or tissue comprising: providing at least one first degradable layer which provides said at least one surface and which is disposed about a central core, and at least one second degradable layer between said first degradable layer and the central core, wherein said first degradable layer has a first degradation rate and said second degradable layer has a second degradation rate such that said at least one first degradable layer degrades more rapidly than said at least one second degradable layer on contact with bodily fluid or tissue.
- FIG. 1 is a schematic representation of an implantable device having nano-craters on the surface of the device.
- FIG. 2 is a Schematic diagram of a process to form nano-craters in a surface using a mask and etching techniques
- FIG. 3 is a schematic representation of an implantable device having two degradable layers.
- FIG. 4 illustrates some of the results of the number of cells correlated to pore size in a PLLA polymer.
- FIG. 5 also illustrates some of the results of the number of cells correlated to pore size in a PLGA polymer sample.
- FIG. 6 illustrates results correlating inter-pore distance to cell attachment and growth of the endothelial cells.
- the reduced thrombogenicity of an implantable device is achieved by enhancing and/or promoting endothelialization of the surface of the implantable device that contacts bodily fluid or tissue.
- This aspect of the invention is based on the surprising discovery that the inclusion of nano-craters on a surface of an implantable device that is intended to come into contact with bodily fluids or tissues, such as blood, advantageously improves endothelial cell attachment to the surface.
- the inclusion of the nano-craters therefore assists in the propagation of endothelial cells on the surface of the device. It is believed that the improved attachment and propagation of endothelial cells on the surface is a result of the nano-craters on the surface acting as foci for endothelial cell attachment.
- This aspect of the invention is particularly suited for manufacture of implantable devices that are intended to be in long-term contact with bodily fluids or tissues, particularly in long-term contact with blood.
- the reduced thrombogenicity is achieved by providing a surface layer that degrades in a controlled fashion, such that any thrombus that is formed at the surface is removed as the surface layer degrades.
- This aspect of the invention is based on the discovery that by providing the surface with layers having different degradation rates, it is possible to remove any thrombus formed on the surface in a controlled fashion, by degradation of each successive layer.
- This aspect of the invention is particularly suited for manufacture of implantable devices that are intended to be in short-term contact with bodily fluids or tissues, particularly blood.
- the implantable device described herein may be any device that would benefit from the reduced thrombogenicity of a surface, including by enhancement of the endothelialization of a surface or by degradation of surface that comes in contact with bodily fluid or tissue, as described below, so as to reduce or remove thrombus formation on such a surface, particularly where such a surface is a blood-contacting surface, when the device is in use.
- the device is useful not only for in vivo applications, but also in vitro applications.
- the device is not particularly limited, but should be considered to include any device that is intended for contact with bodily fluids or tissues, particularly blood, including conduits, grafts, valves, dialysis tubing and stents.
- bodily fluids or tissues includes biologically derived fluids and tissues as well as synthetic substitutes, for example artificial blood.
- endothelialization refers to the growth and/or proliferation of endothelial cells on a surface, such as the blood-contacting surface, or an implantable device. Promoting or enhancing endothelialization of a surface refers to promoting, enhancing, facilitation or increasing the attachment of, and growth of, endothelial cells on the surface.
- the surface of a device for implantation into a subject is preferably biocompatible.
- biocompatible means that a substance is minimally toxic or irritating to biological tissue, such as to be sufficiently tolerated in the body without adverse effect.
- the surface may be formed of a material, which is different from the material that forms the surface and which is used as a support. Alternatively, the device and surface may be formed of the same material.
- Suitable materials for forming the surface include biostable polymers, for example, polyethylene, polyurethane, polyolefin, or polyethylene terephthalate and degradable polymers, including degradable by chemical means or by exposure to radiation, for example, poly-lactide (PLA) including poly-L-lactide (PLLA), poly-glycolide (PGA), poly(lactide-co-glycolide) (PLGA) or polycaprolactone.
- PLA poly-lactide
- PLLA poly-L-lactide
- PGA poly-glycolide
- PLGA poly(lactide-co-glycolide)
- the degradable polymer may be biodegradable, meaning that the substance will readily degrade in an environment that is, or that is equivalent to, the body of a subject, for example when in contact with bodily fluid or tissue.
- suitable materials that can be used to form an implantable device, or to provide the surface of an implantable device, are generally known in the art and examples of such materials are outlined in U.S. Pat. No. 5,744,515, which is herein incorporated by reference.
- preferred materials include synthetic polymers, including oligomers, homopolymers, and copolymers resulting from either addition or condensation polymerization.
- suitable addition polymers include, but are not limited to, acrylics such as those polymerized from methyl cerylate, methyl methacrylate, acrylic acid, methacrylic acid, acrylamide, hydroxyethyl acrylate, hydroxyethyl methacrylate, glyceryl scrylate, glyceryl methacrylate, methacrylamide and methacrylamide; vinyls such as styrene, vinyl chloride, binaly pyrrolidone, polyvinyl alcohol, and vinyls acetate; polymers formed of ethylene, propylene, and tetrafluoroethylene.
- acrylics such as those polymerized from methyl cerylate, methyl methacrylate, acrylic acid, methacrylic acid, acrylamide, hydroxyethyl acrylate, hydroxyethyl methacrylate, glyceryl scrylate, glyceryl methacrylate, methacrylamide and methacrylamide
- vinyls such as styrene
- condensation polymers include, but are not limited to, nylons such as polycaprolactam, polylauryl lactam, polyhexamethylene adipamide, and polyhexamethylene dodecanediamide, and also polyurethanes, polycarbonates, polyamides, polysulfones, poly(ethylene terephthalate), polylactic acid, polyglycolic acid, polydimethylsiloxanes, and polyetherketones.
- Suitable materials include metals and ceramics.
- the materials include, but are not limited to, nickel, titanium, nickel-titanium alloys such as Nitinol, stainless steel, cobalt and chromium.
- the ceramics include, but are not limited to, silicon nitride, silicon carbide, zirconia, and alumina, as well as glass and silica, ePTFE (Expanded polytetrafluoroethylene) is a preferred substrate material for use in fabricating implantable devices of the present invention, and particularly for fabricating vascular grafts.
- Suitable ePTFE is available in the form of vascular grafts from such sources as IMPRA, Inc., Tempe, Ariz. Commercially available grafts are constructed of ePTFE and supplied in sterile form in a variety of configurations, including straight, tapered and stepped configurations.
- device 100 is stent, with an exterior surface 102 a and an interior surface 102 b which lines the lumen of the stent, both of which have reduced thrombogenicity meaning that they have a reduced tendency to promote, induce or facilitate formation of thrombi when in contact with bodily fluid or tissue.
- thrombogenicity meaning that they have a reduced tendency to promote, induce or facilitate formation of thrombi when in contact with bodily fluid or tissue.
- surface 102 b contacts blood, including platelets, it is particularly important that surface 102 b be rendered less thrombogenic, as described herein.
- Device 100 may comprise a polymeric stent fabricated as disclosed in U.S. patent application Ser. No. 10/867,617 filed Jun. 15, 2004 (U.S. Pat. Pub. 2005/0021131 A1), which is incorporated herein by reference in its entirety.
- the stent as shown and described, may comprise a polymer that is at least partially amorphous and which undergoes a transition from a pliable, elastic state at a first higher temperature to a brittle glass-like state at a second lower temperature as it transitions through a particular glass transition temperature.
- This particular stent may be comprised of at least a first layer and a second layer where the first layer includes a first polymer that is at least partially amorphous and a second layer that is also at least partially amorphous.
- the stent may be formed to have a first shape at a relatively lower temperature and a second shape at a relatively higher temperature.
- the inner and/or outer layer of the stent 100 may be processed to have nano-crater 104 as described herein.
- a substantially uniform layer of nano-craters 104 are distributed on surface 102 a and 102 b , meaning that nano-craters 104 of substantially similar depth are distributed on the surfaces 102 a and 102 b to form a discernible layer having such nano-craters. It has advantageously been found that the provision of such nano-craters 104 enhances endothelialization of surface 102 a and 102 b , resulting in reduced thrombogenicity.
- the stent 100 is suitable for long-term implantation in the body of a subject.
- nano-crater means indentations or depressions provided on a surface. Generally the indentations are on the nanometer scale. In different embodiments, the nano-craters have an average diameter of between about 30 nm and about 150 nm.
- the stent 100 has nano-craters 104 sufficiently distributed over surfaces 102 a and 102 b to promote or enhance endothelialization, preferably over the entirety of surface 102 a and 102 b .
- the nano-craters 104 may be regularly or irregularly distributed over surfaces 102 a and 102 b . In some embodiments, adjacent craters may be spaced about 200 nm or greater apart.
- Such nano-craters 104 may be suitably shaped, having a regular or irregular shape, provide that endothelialization of the surfaces 102 a and 102 b having the nano-craters 104 is enhanced and/or promoted.
- the nano-craters 104 may be hemi-spherical, hemi-cylindrical or elliptical.
- the size and shape of the nano-craters 104 can be controlled to provide a unique surface morphology. By varying this surface morphology, the range of sizes that selectively promote endothelial cell attachment while not being reception to platelet attachment, can be readily ascertained.
- surfaces 102 a and 102 b of the stent 100 can be chemically modified so as to further enhance or promote endothelialization, for example when implanted in a subject's body.
- endothelial cells migrate into the device from tissue that is external to (usually above or below) the implanted device.
- the device itself must be sufficiently porous to permit the endothelial cells to migrate into it.
- a coronary stent such as the Palmaz stent (U.S. Pat. No. 6,379,383) is an example of such a device.
- This type of endothelialization may be achieved by coating an implantable device with a radiation-sensitive bioerodible polymer followed by irradiation with an electron beam to generate the nano-craters, as it set out below.
- the second method of endothelialization involves migration of endothelial cells longitudinally into the device (e.g., in the lumen of a stent implanted in a blood vessel) from tissue adjacent to the device.
- porosity of the implantable device is not required, as endothelial cell attachment occurs from within a lumen or cavity of the device.
- the number of endothelial cells that are capable of this type of attachment is lower than those that can be achieved by transmural endothelialization.
- nano-cratered surfaces will enhance selective endothelial cell attachment on non-porous devices
- the production and attachment of these endothelial cells in vivo may be enhanced using certain growth-stimulating molecules and adhesion-promoting molecules.
- growth-stimulating molecule refers to a molecule that stimulates or induces the differentiation, growth and proliferation of endothelial cells.
- Growth-stimulating molecules include peptides, proteins and glycoproteins, including hormones, capable of inducing an endothelial cell to grow and divide.
- Adhesion-promoting molecule refers to a molecule that promotes or encourages adhesion or attachment of an endothelial cell to a surface.
- Adhesion-promoting molecules include peptides, proteins and glycoproteins capable of binding a cell to a substrate or to an adjacent cell.
- surfaces 102 a and 102 b of the stent 100 include growth-stimulating molecules and/or adhesion-promoting molecules dispersed therein, which facilitate enhanced production of endothelial cells and their attachment to the nano-cratered surfaces 102 a and 102 b.
- Suitable growth-stimulating molecules include granulocyte colony stimulating factor (gCSF), platelet-derived endothelial cell growth factor (PD-ECGF), fibroblast-derived endothelial cell growth factor alpha, endothelial cell growth factor beta, endothelial cell growth factor 2a and endothelial call growth factor 2b.
- GCF granulocyte colony stimulating factor
- PD-ECGF platelet-derived endothelial cell growth factor
- fibroblast-derived endothelial cell growth factor alpha alpha
- endothelial cell growth factor beta endothelial cell growth factor 2a
- endothelial call growth factor 2b endothelial call growth factor
- adhesion molecules are described in U.S. Pat. No. 5,774,515, which is herein incorporated by references. They are typically large, naturally occurring proteins or carbohydrates, with molecular weights above 100,000 daltons. In vivo, adhesion molecules are typically able to bind to specific cell surface receptors, and mechanically attached cells to the substrate or to adjacent cells. In addition to promoting cell attachment, suitable adhesion molecules can promote other cell responses including cell migration and cell differentiation (which in turn can include the formation of capillary tubes by endothelial cells).
- Preferred adhesion molecules include substrate adhesion molecules (SAM's) such as the proteins laminin, fibronectin, collagen, vitronectin, and tenascin, and adhesion peptides or functional synthetic analogs derived from SAM's.
- SAM's substrate adhesion molecules
- Other suitable adhesion molecules include cell-to-cell adhesion molecules (CAM's) such as N-cadherin and P-cadherin.
- Parent adhesion proteins typically have one or more active peptide domains that bind to cell surface receptors and which domains produce the cell attachment, migration, and differentiation effects of the parent adhesion proteins. These domains consist of specific amino acid sequences, several of which have been synthesized and reported to promote the adhesion of endothelial cells. These domains and functional analogs of these domains are termed “adhesion peptides”.
- adhesion molecules are adhesion peptides and desirably, adhesion peptides have about 3 to about 30 amino acid residues in their amino acid sequences.
- Adhesion peptides from fibronectin include, but are not limited to, RGD (arg-gly-asp) [SEQ ID NO.:1], REDV (arg-glu-asp-val) [SEQ ID NO.:2], and C/H—V (WQPPRARI or trp-gln-pro-pro-arg-ala-arg-ile) [SEQ ID NO.:3].
- Adhesion peptides from laminin include, but are not limited to, YIGSR (tyr-ile-gly-ser-arg) [SEQ ID NO.:4] and SIKVAV (ser-ile-lys-val-ala-val) [SEQ ID NO.:5] and F-9 (RYVVLPRPVCFEKGMNYTVR or arg-tyr-val-val-leu-pro-arg-pro-val-cys-phe-glu-lys-gly-met-asn-tyr-thr-val-arg) [SEQ ID NO.:6].
- Adhesion peptides from type IV collagen include, but are not limited to, Hep-III (GEFYFDLRLKGDK or gly-glu-phe-tyr-phe-asp-leu-arg-leu-lys-gly-asp-lys) [SEQ ID NO.:7].
- an anti-thrombotic molecule may be included on the surfaces 102 a and 102 b of the stent 100 by any suitable means, in amounts sufficient to minimize any platelet attachment during the process of endothelialization.
- an “anti-thrombotic molecule” is a molecule that reduces or prevents the formation of thrombi or clots on the surface of an implantable device that contacts bodily fluid or tissue, including when implanted in a subject's body.
- Anti-thrombotic molecules include, without limitation, heparin, and small molecules, such as benzamidine compounds, bicyclic pyrimidine compounds, nitro compounds, thio acid compounds, and proteins and peptides, including tissue-type plaminogen activator (t-PA), protein S and protein C.
- the implantable device may be formed entirely from a single material and standard methods know in the art may be used to fashion the device. For example, a mold may be used, and a liquid polymer may be poured into the mold. This methods used will depend on the particular material used and the particular medical device that is to be formed.
- the device may be formed by rolling a sheet or film of material, or by winding a thin strip of material into a helix, as is known in the art.
- the nano-craters may be readily formed on each side of the sheet or strip, as discussed below, prior to rolling or winding to form the stent.
- the implantable device may also be formed from a substrate material and another material applied to the substrate material to form a bodily fluid or tissue contracting surface by any suitable means, for example, by spin-coating from a solution or suspension, and the nano-craters are subsequently introduced into the surface.
- This surface layer should have sufficient thickness to introduce nano-craters having depth sufficient to enhance or promote endothelialization.
- the following illustration of two possible approaches for forming the nano-craters 104 are provided.
- the nano-craters 104 may be introduced through controlled degradation of the surfaces 102 a and 102 b of the stent 100 , as depicted in FIG. 2 .
- discrete portions of surfaces 102 a and 102 b are etched using a degradative process, for example, by exposing the polymer surface to electron beam radiation or by treating with a chemical that will degrade the surface, for example, strong alkali.
- a higher density material for example a silicon-based polymer or an acrylic polymer, may be patterned over surface 102 a and 102 b in which the nano-craters 104 are to be introduced, in a pattern that defines the desired distribution and depth of the nano-craters.
- a focused ion beam may be used to form the desired pattern in the mask material which is layered on the degradable surface 102 a and 102 b.
- the surface material in the degraded areas may then be leached out using water or solvent in which the degraded portions of the surface material are soluble, but which will not dissolve the non-degraded regions of the surface.
- the mask material may then be subsequently removed, for example by dissolution in a suitable solvent that dissolves the mask material but not the polymer surface 102 a and 102 b.
- PLGA, PLLA, PGA, polycaprolactone or polyethylene may be employed to form the stent 100 or surfaces 102 a and 102 b of the stent 100 , both of which degrade in the dry state under electron-beam irradiation.
- the degree of degradation may be controlled using the well-known effects of attenuation with depths of an incident electron beam.
- the depth of penetration of the incident electron beam is generally proportional to the electron energy or the accelerating voltage being used.
- This depth-dose distribution is determined by the absorption mechanism of mono-energetic electron beams having electron energy, eV, for a material of density p.
- the higher the density of a given material the grater the attenuation effect on the electron beam.
- This attenuation effect will result in a varying radiation dose across the thickness of the surface and patterned higher density material, resulting in a variation of molecular weight of the polymer across the thickness of the surface.
- An example of utilizing an incident electron beam for patterning a surface of a polymeric sample may include use of electron beam lithography, which is typically used in the semiconductor electronics industry for patterning integrated circuits and biosensors.
- a polymeric substrate having a radiation-sensitive film or resist may be placed in a vacuum chamber of a scanning-electron microscope and exposed by an electron beam under digital control. Because the beam width may be adjusted to range from a few picometers to several nanometers, an etched pattern may be formed by the beam across the polymer surface.
- the size and shape of the nano-craters 104 may be accurately controlled by this method, for example by controlling the does of the radiation, and the density of the material used to mask, as well as the pattern in which the masking material is applied. This results in a unique surface morphology, as discussed above, that selectively promotes endothelial cell attachment, while not being receptive to platelet attachment.
- chemical means can be used with the above-described masking method to produce nano-craters at the surfaces 102 a and 102 b .
- sodium hydroxide may be used to dissolve PLA in regions that are not protected by the alkali-resistant mask material, and the dissolved material may then be rinsed away in water to form nano-craters 104 .
- the mask may be removed as described above.
- the nano-craters 104 may alternatively be formed on the surfaces 102 a and 102 b of the stent 100 by including nano-particles that are leachable from the surfaces 102 a and 102 b.
- nano-particle is any granular or particulate material in which the particulates have dimensions in the nano-meter range.
- the nano-particles may be irregularly shaped, or may be of well-defined size and shape, and may be leached from the surface leaving behind nano-craters corresponding to the size and shape of the nano-particles.
- the nano-particles may be formed of any granular or particulate material which can be embedded in the material used to form surface 102 a and 102 b , which will not dissolve in or become irreversibly bound to the material, and which can then be subsequently leached from the material.
- the nano-particles can be formed from an inorganic salt, such as sodium chloride, form gelatin, sugar, chitosan, or polyvinyl pyrrolidone.
- the nano-particles may be suspended in a dilute solution of a polymer being used to form the implantable device or more preferably, the surface of the implantable device which may then be spin-coated onto the substrate of the device at a desired thickness.
- the thickness will usually be in the micrometer range.
- these particles on the surface are either leached out upon exposure to water or another suitable solvent, or are eroded once the device comes in contact with bodily fluid or tissue, for example when stent 100 is implanted, leaving behind a surface with well defined nano-craters 104 of know dimensions.
- the dimensions of the nano-craters 104 may be varied by varying the size and shape of the nano-particles dispersed in the polymer.
- the nano-craters may be created, for example by irradiation, and concurrently the surface may be modified to release adhesion-promoting molecules and/or growth-stimulating molecules, for example into a lumen or cavity of the implantable device.
- the adhesion-promoting molecules and/or growth-stimulating molecules may be passed to a polymer used to form the implantable device or the surface of the implantable device prior to coating the polymer on the substrate of the implantable device, and forming nano-craters.
- adhesion-promoting molecules and growth-stimulating molecules may typically be proteins, which are sensitive biomolecules that may be denatured by addition to a liquid polymer, or when subjected to high intensity radiation.
- the adhesion-promoting molecules and/or growth-stimulating molecules may first be encapsulated in nano-particles of well-defined size and shape as it known in the art, for example, as described in U.S. Pat. No. 6,589,562 which is herein fully incorporated by reference.
- the nano-particles may be leached out as discussed above, leaving behind the nano-craters and simultaneously releasing the adhesion-promoting molecules and/or growth-stimulating molecules, for example into a lumen.
- the nano-particles, when containing adhesion-promoting molecules and/or growth-stimulating molecules for delivery to bodily fluid or tissue comprise a material that is soluble in bodily fluid or tissue, for example, gelatin.
- An anti-thrombotic molecule may be included in the nano-crated surface of an implantable device in a similar manner.
- an implantable device with reduced thrombogenicity is achieved by providing the device with a surface that will degrade in a layered fashion when it contacts bodily fluid or tissue.
- This embodiment is useful for applications in which the device will be in contact with bodily fluid or tissue for a relatively short period of time, for example, a catheter or dialysis tubing that is in such contact for less than 24 hours.
- the layers degrade relatively quickly, so as to prevent the formation of thrombi. This means that the degradation time for a given layer upon contacting bodily fluid or tissue may be, for example, between about 5 minutes and about 1 hour.
- a stent 100 ′ has first degradable layers 106 a and 106 b disposed about a central core 110 , and which layers provide surfaces 102 ′ a and 102 ′ b that comes into contact with bodily fluid or tissue, including blood, and second degradable layers 108 a and 108 b , between layers 106 a and 106 b , respectively, and the central core 110 of stent 100 ′.
- the stent 100 ′ has a first surface 102 ′ a , which forms the exterior surface of the stent and an interior surface 102 ′ b which defines the lumen of the stent.
- the second degradable layers 108 a and 108 b are the inner layer relative to the outer surfaces 102 a ′ and 102 ′ b , respectively, and have a slower degradation rate than the first degradable layers 106 a and 106 b . Therefore, on contact with bodily fluid or tissue, there is a peeling effect resulting from successive degradation of first degradable layers 106 a and 106 b followed by degradation of the second degradable layers 108 a and 108 b , and any thrombus formation on surface 102 ′ a and 102 ′ b is removed as the layers erode.
- the stent 100 ′ may also be configured and comprised in the manner as shown and described in U.S. patent application Ser. No. 10/867,617, which has been incorporated above by reference in its entirety.
- the stent 100 ′ configured as disclosed in U.S. patent application Ser. No. 10/867,617 may comprise the central core 110 having the multiple degradable layers disposed thereon. In other variations, it may be possible to have the multiple degradable layers correspond to the multiple layers comprising the stent structure.
- the degradable layers 106 a and 106 b and 108 a and 108 b may be formed from any biodegradable polymers that are generally known in the art and described above and hereafter.
- suitable polymers include polylactic acid (PLA) and polyglycolic acid (PGA) and copolymers of PLA and PGA (PLGA). These polymers may be amorphous or semi-crystalline.
- layers 106 a and 106 b may comprise PLA and the layers 108 a and 108 b may comprise PLGA, particularly PLDA 80/20; PLGA 75/25; or PLGA 53/47, wherein the numbers in the copolymer represent the percentage of PLA and PGA by weight, respectively, included in the copolymer.
- each layer 106 a and 106 b and 108 a and 108 b is in the micrometer or sub-micrometer range, for example about 0.5 ⁇ m to about 10 ⁇ m.
- the central core 110 may comprise a different material than layers 106 a , 106 b , 108 a and 108 b , and the material comprising the respective layers may be applied to central core 110 .
- stent 100 ′ may be formed of a single polymeric material but having first and second degradable layers of different average molecular weights of the polymer than found in central core 110 , so as to form the discrete layer 106 a , 106 b , 108 a , and 108 b about central core 110 , as described below.
- the degradable layers 106 a and 106 b and 108 a and 108 b having varying degradation rates are provided on the central core 110 .
- Polymers having different degradation rates can be selected and applied successively such that the layers 108 a and 108 b comprise a polymer with a slower degradation rate.
- a polymer with a faster degradation rate is selected for layers 106 a and 106 b such that layer 106 a and 106 b degrade more rapidly and remove any thrombus that may have formed on the surfaces 102 ′ a and 102 ′ b , respectively.
- a layered device having first and second degradable layers may comprise additional degradable layers, and that the degradation rate of each degradable layer increases with each successively inward layer such that the outer-most layer degrades more quickly and that the inner-most layer degrades most slowly.
- a layered device may comprise the following layers disposed about a central core: PLA; PLGA 80/20; PLGA 75/25; and PLGA 53/47 in the given order with PLGA 53/47 being the outer-most layer.
- the suitable number of layers to be applied can be readily determined and will depend on the degradation rates of the layers and the particular type of device and its intended use, including the intended duration of contact with bodily fluid or tissue.
- Each of such layers may be spin-coated or solvent cast on to a substrate material forming the implantable device, using a solution or suspension containing, for example, about 10 to about 40% polymer by weight.
- a solution or suspension containing, for example, about 10 to about 40% polymer by weight.
- other suitable means of applying thin layers of a polymer to a substrate may also be employed, for example, vapour deposition.
- controlled degradation of a surface of an implantable device may be effected, for example, using radiation such as electron beam radiation.
- radiation such as electron beam radiation.
- This method utilizes the attenuation effect of electron beam radiation within an irradiated material.
- a single biodegradable material may be applied to the surface of an implantable device as described above and then irradiated to provide layers having different average molecular weights of the biodegradable material, and therefore varying degradation rates.
- the suitable thickness of the material to be applied will typically be in the micrometer range, for example about 1 micron to about 20 microns, and can be readily determined. The desired thickness will depend on the particular polymer used and on the particular type of device and its intended use, including the intended duration of contact with bodily fluid and tissue.
- the mechanism of attenuation can be described as the loss of energy of the accelerating electrons.
- the depth of penetration is proportional to the electron energy or the accelerating voltage, and is attenuated in a manner proportional to the density of the material being penetrated.
- This attenuation effect will result in a varying radiation does through the depth of the material as the beam is attenuated as it travels deeper into the material, with the exterior surface receiving the strongest does of radiation. This will result in a variation of molecular weight in the surface material as a function of penetration depth or material thickness.
- the above-described devices can provide an implantable device having reduced thrombogenicity on contact with bodily fluid or tissue, for example when implanted, as compared to that typically observed with implantable medical devices.
- Standard surgical methods for implanting medical devices are known in the art. The method of implantation and duration of implantation will depend on the type of implantable device used, for example, a stent or a valve, the purpose of implantation and the disorder or condition that is to be treated with the implantable medical device.
- a method for reducing thrombogenicity, and for enhancing or promoting endothelialization, of an implantable device having at least one surface for contacting bodily fluid or tissue is contemplated.
- the method comprises providing on the at least one surface a plurality of nano-craters that enhance or promote endothelialization of the at least one surface.
- the method comprises providing at least one first degradable layer which provides said at least one surface and which is disposed about a central core, and at least one second degradable layer between said first degradable layer and the central core, wherein said first degradable layer has a first degradation rate and said second degradable layer has a second degradation rate such that said at least one first degradable layer degrades more rapidly then said at least one second degradable layer so as to remove any thrombus that may be formed on said at least one surface.
- each polymer was first dissolved in chloroform. Nano-sized salt particles were ground and sieved, and then dispersed in the polymer solution with constant stirring until the particles were visually uniformly dispersed. The polymer concentration was chosen such that it had sufficiently high viscosity to maintain a stable dispersion. The dispersion was then cast as a film of required thickness using a coater. The film was dried in an oven at 37° C., and then left at room temperature for several days in a dry environment. The dried films were immersed in water for 14 days, with constant exchange of the water. The salt nano-particles were thus leached out, and the resulting film was dried again at 37° C. and at room temperature.
- Control films were prepared as pure polymer films without any surface modification.
- PLLA and PLGL films having nano-craters in the surface were obtained by leaching out incorporated nano-particles of NaCl, as indicated in Table 1.
- porogen leaching of surfaces may be utilized to yield a surface which enhances endothelial cell growth over a defined range of surface features.
- surface pores were created by filling polymers such as Poly caprolactone (PCL), Poly L-lactide (PLLA), Poly (lactide-co-glycolide), etc. (although any of the other suitable polymers described herein may be utilized) with leaching agents of sugar and gelatin.
- the sugar and gelatin particles ranged in size from 20 to 90 microns in diameter (although particles as small as 5 microns may also be utilized) where the average particle sizes typically ranged from 20, 45, and 90 microns.
- the leaching agents were added in concentrations ranging from 1 to 10% by weight in the polymer. More particularly, the leaching agents were added in concentrations ranging from 1%, 5%, and 10% by weight in the polymer.
- the leaching agents were then leached out with water from the polymer for a period of 10 to 12 days and the surface porosity was characterized by a scanning electron microscope (SEM) for crater dimensions and inter-crater spacing. With the physical characteristics determined, the surfaces of the polymer were then exposed to endothelial cells over an 11 day period, at the end of which the cells attached to the surface were counted and correlated to the surface features.
- SEM scanning electron microscope
- FIG. 4 illustrates some of the results of the number of cells correlated to pore size in a PLLA polymer sample at day 9, which is representative of the results.
- FIG. 5 also illustrates some of the results of the number of cells correlated to pore size in a PLGA polymer sample (specifically PLGA 80/20) also at day 9.
- each sample was prepared utilizing the methods described above.
- endothelial cell growth appeared better on PLGA 80/20 samples than on PLLA samples.
- both gelatin and sugar pyrogens appear to act similarly and regardless of the porogen used, cell growth appears inversely dependent on pore size.
- gelatin appeared to be optimal for use as a porogen in the size range of about 5 to 40 microns at concentrations of about 5 to 10% in the starting solution.
- the PCL samples also prepared as described above, showed growth of endothelial cells although the growth did not appear dependent on pore size in the range studied.
- endothelial cell attachment and proliferation is higher at lower crater sizes (between about 5-10 microns) and decreases with higher crater size up to about 90 microns; however, compared to controls (no craters), all the samples showed enhanced endothelial cell attachment.
- the inter-pore distances along the polymer surfaces were varied from an average of about 50 microns to 250 microns. As illustrated by the results in FIG. 6 , an inter-pore distance ranging from about 50 to 100 microns and more preferably between 50 to 80 microns appeared optimal for attachment and growth of the endothelial cells.
- endothelial cell growth appears to correlate inversely to pore size on surfaces of PLLA and PLGA samples, but not to PCL samples.
- pore size is decreased (e.g., down to about 5 to 10 microns)
- endothelial cell growth is increased.
- PCL showed good endothelial cell growth on its surface.
- chemicals such as sodium hydroxide may be used to dissolve PLA in regions unprotected by an alkali-resistant mask material where the dissolved material may be rinsed away in water to form nano-craters.
- the polymer surface may be first irradiated prior to etching with the sodium hydroxide to enhance the etching process.
- samples of PLGA, PCL, and PLLA were first irradiated with an electron beam and then etched using the sodium hydroxide, as described above, for a period of 16 hours to create surface features.
- the average surface roughness of the samples was measured using an atomic force microscope (AFM) and the etched samples were then exposed to endothelial cells. Growth was quantified over a period of 15 days and the irradiated and etched samples were compared to control samples after 4 days, 8 days, and 15 days.
- Table 2 shows a comparison of the results for sample roughness between the irradiated and control samples where the MTS value is an indication of the number of active cells.
- irradiating samples prior to etching with sodium hydroxide gives surface features that are rougher than control samples.
- Table 3 shows a comparison of the results for the irradiated and control samples with respect to live cell growth and total cell growth.
- the surface-modified samples show enhanced endothelial cell growth for PLGA and PCL samples except for PLLA samples.
- the endothelial cell growth also appeared to correlate well with overall surface roughness of PLGA and PCL samples where endothelial cell growth increases as surface roughness increases.
Abstract
Description
TABLE 1 |
Results of Endothelialization of Nano-Cratered Surfaces. |
First | ||||||
endothrlial | ||||||
Sample | Surface | Cell Seeding | call | Result at days | ||
Material | Preparation | Treatment | (cells/sq cm) | |
4/5 | |
Control PLLA | Polymer + | |
20000 | 36 | Day | 5 |
Solvent PLLA | approximately | |||||
20% | ||||||
confluency | ||||||
Contold | Polymer + | NIL | As Above | 36 | Day | 5 |
PLGA | Solvent PLGA | approximately | ||||
80:20 | 40% | |||||
confluency | ||||||
PLLA with | Polymer + | Leached NaCl | As Above | 2 hours | At |
|
Nanocraters | Solvent PLLA | 99% purity | about 70% | |||
<90 Microns | confluency | |||||
1% | Seen. | |||||
concentration | ||||||
Leaching | ||||||
period 15 | ||||||
days. | ||||||
PLGA with | Polymer + | Leached NaCl | As Above | 6 hours | At |
|
nanocraters | Solvent PLGA | 990% Purity | about 5% | |||
80:20 | <90 microns | confluency | ||||
1% | Seen. | |||||
concentration | ||||||
Leaching | ||||||
period 15 | ||||||
days. | ||||||
TABLE 2 |
Results of Comparison For Irradiated and Control Samples With Respect to |
Sample Roughness and Cell Growth. |
AFM Avg surface | MTS Average | MTS Average | MTS Average | ||
Roughness (Scan | Absorbance after 4 | Absorbance after 8 | Absorbance after 15 | ||
Size 50 μm) | Static Contact Angle | days | days | days | |
PLGA Control | 3.3 ± 1 | 7.32 ± 1 | 0.51 | 0.37 | 0.26 |
PLGA Modified | 93 ± 3 | 57.4 ± 2 | 0.57 | 0.29 | 0.45 |
PLLA Control | 646 ± 9 | 94.2 ± 2 | 0.40 | 0.29 | 0.40 |
PLLA Modified | 333 ± 27 | 63.4 ± 1 | 0.51 | 0.17 | 0.27 |
PCL Control | 259 ± 20 | 80.2 ± 3 | 0.39 | 0.24 | 0.28 |
PCL Modified | 390 ± 16 | 61.8 ± 1 | 0.53 | 0.38 | 0.39 |
*Modified = Ebeam with 2.5 Mrads + 16 hours 0.1N NaOH immersion |
TABLE 3 |
Results of Comparison For Irradiated and Control Samples With Respect to |
Live Cell Growth and Total Cell Growth. |
Hemocytometer | Hemocytometer | Hemocytometer | Hemocytometer | Hemocytometer | Hemocytometer | |
Avg Live Cells | Avg total Cells | Avg Live Cells | Avg total Cells | Avg Live Cells | Avg total Cells | |
Count after 4 day | Count after 4 day | Count after 8 day | Count after 8 day | Count after 15 day | Count after 15 day | |
PLGA Control | 5400 | 9600 | 9300 | 15800 | 9400 | 22700 |
PLGA Modified | 8800 | 12800 | 9700 | 17800 | 17800 | 31400 |
PLLA Control | 5300 | 9000 | 3800 | 4900 | 13500 | 27700 |
PLLA Modified | 6500 | 8800 | 3300 | 4800 | 13600 | 27400 |
PCL Control | 4400 | 9200 | 1530 | 3800 | 3060 | 12000 |
PCL Modified | 4100 | 7600 | 9830 | 14200 | 5300 | 23400 |
*Modified = Ebeam with 2.5 Mrads + 16 hours 0.1N NaOH immersion |
Claims (6)
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